Cyanotypic frequencies in adjacent and mixed populations of Trifolium occidentale Coombe and Trifolium repens L. are regulated by different mechanisms

The cyanogenic polymorphism in Trifolium repens is caused by the variation in two genes, the interaction of which produces four distinct cyanotypes. Along the Atlantic coasts of Bretagne, T. repens is sometimes found in populations mixed with the related species Trifolium occidentale, although the latter species usually occurs only in a narrow fringe along the coast, whereas T. repens is a more inland species. No plants of T. occidentale have ever been reported to have linamarase activity. Indeed, of 763 T. occidentale plants studied, none contained linamarase activity. However, the variation in the proportion of cyanotypes in T. repens was enormous, even between sites less than 2km apart. Our results confirm the presumption that T. repens and T. occidentale are indeed separate species. Both the fact that T. occidentale plants never contain linamarase activity, and the difference in proportion of plants with cyanoglucosides in mixed stands show that gene flow between the species must be rare. These dissimilar distributions strongly indicate that cyanotypic frequencies in adjacent and mixed populations of the very closely related species T. occidentale and T. repens are regulated by different mechanisms     

Decreased anthocyanidin reductase expression strongly decreases silver birch (Betula pendula) growth and alters accumulation of phenolics

Phenolics, formed via a complex phenylpropanoid pathway, are important defensive agents in plants and are strongly affected by nitrogen (N) fertilization. Proanthocyanidins (PAs) are one possible endpoint of the phenylpropanoid pathway, and anthocyanidin reductase (ANR) represents a key enzyme in PA biosynthesis. In this study, the expression of silver birch (Betula pendula) anthocyanidin reductase BpANR was inhibited using the RNA interference (RNAi) method, in three consequent BpANR RNAi (ANRi birches) lines. The growth, the metabolites of the phenylpropanoid pathway, and the number of resin glands of the ANRi birches were studied when grown at two N levels. ANRi birches showed decreased growth and reduction in PA content, while the accumulation of total phenolics in both stems and leaves increased. Moreover, ANRi birches produced more resin glands than did wild‐type (WT) birches. The response of ANRi birches to N depletion varied compared with that of WT birches, and in particular, the concentrations of some phenolics in stems increased in WT birches and decreased in ANRi birches. Because the inhibition of PAs biosynthesis via ANR seriously affected birch growth and resulted in accumulation of the precursors, the native level of PAs in plant tissues is assumed to be the prerequisite for normal plant growth. This draws attention to the real plant developmental importance of PAs in plant tissues.     

Ancestors of white clover ( Trifolium repens L.), as revealed by isozyme polymorphisms

Isozymes were used to study the putative ancestors of white clover ( Trifolium repens L.). Ten enzymes were examined, and 18 loci were resolved via starch-gel electrophoresis for accessions representing T. repens, Trifolium isthmocarpum, Trifolium nigrescens, Trifolium occidentale and Trifolium uniflorum, in addition to two more distantly related species, Trifolium alpinum and Trifolium purseglovei. Nei's genetic identities indicate that T. uniflorum and T. nigrescens are the closest relatives of T. repens. The isozyme data thus support a hypothesis that the two genomes of the tetraploid T. repens could have been derived from hybridization between T. nigrescens and T. uniflorum. This conclusion is further supported by shared alleles between T. repens, T. nigrescens and T. uniflorum. However, the origin of T. repens is somewhat obscured by the presence of shared alleles between T. repens and both T. occidentale and T. isthmocarpum, suggesting that introgression of genes from the latter two species into T. repens may also have taken place. High values of genetic identity are shared between T. occidentale with T. nigrescens and T. uniflorum, also indicative of introgression. Alternatively the presence of shared alleles among the five species may reflect their recent common ancestry.     

An integrated approach to demonstrating the ANR pathway of proanthocyanidin biosynthesis in plants

Proanthocyanidins (PAs) are oligomers or polymers of plant flavan-3-ols and are important to plant adaptation in extreme environmental conditions. The characterization of anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) has demonstrated the different biogenesis of four stereo-configurations of flavan-3-ols. It is important to understand whether ANR and the ANR pathway widely occur in the plant kingdom. Here, we report an integrated approach to demonstrate the ANR pathway in plants. This includes different methods to extract native ANR from different tissues of eight angiosperm plants (Lotus corniculatus, Desmodium uncinatum, Medicago sativa, Hordeum vulgare, Vitis vinifera, Vitis bellula, Parthenocissus heterophylla, and Cerasus serrulata) and one fern plant (Dryopteris pycnopteroides), a general enzymatic analysis approach to demonstrate the ANR activity, high-performance liquid chromatography-based fingerprinting to demonstrate (-)-epicatechin and other flavan-3-ol molecules, and phytochemical analysis of PAs. Results demonstrate that in addition to leaves of M. sativa, tissues of other eight plants contain an active ANR pathway. Particularly, the leaves, flowers and pods of D. uncinatum, which is a model plant to study LAR and the LAR pathways, are demonstrated to express an active ANR pathway. This finding suggests that the ANR pathway involves PA biosynthesis in D. uncinatum. In addition, a sequence BLAST analysis reveals that ANR homologs have been sequenced in plants from both gymnosperms and angiosperms. These data show that the ANR pathway to PA biosynthesis occurs in both seed and seedless vascular plants.     

Extent of high-affinity iron transport systems in field isolates of rhizobia

A Uruguayan rhizobia collection (67 isolates) obtained from nodules of Medicago sativa, Melilotus albus, Medicago polymorpha, Trifolium subterraneum, Trifolium repens, Trifolium vesiculosum, Lotus corniculatus, Lotus subbiflorus, Lotus pedunculatus, Ornithopus sp. and Adesmia sp. has been examined to assess the occurrence of high affinity iron uptake systems. CAS (Chrome-azurol S)-assay results suggested that most of the free-living form of these microsymbionts may produce siderophores. The highest siderophore production was observed among Medicago and Trifolium microsymbionts whereas no siderophore expression or moderate positive results were found among Lotus microsymbionts; suggesting that microsymbionts of legumes growing on neutral or alkaline soils may express in vitro enhanced siderophore production. Electrophoretic patterns of outer-membrane protein enriched fractions revealed that iron-limited microsymbionts of Medicago sativa, Lotus corniculatus, Lotus subbiflorus, Trifolium repens, Trifolium subterraneum and Ornithopus sp. produced high molecular weight proteins (ranging from 64 to 94 kDa) compared to cells grown in iron-sufficient media.     

白车轴草和紫花苜蓿根瘤的显微及超微结构

通过对白车轴草(Trifolium repens Linn.)和紫花苜蓿(Medicago sativa Linn.)根瘤的显微及超微结构进行观察,发现其根瘤显微结构都由4部分组成,由外向内依次为：保护层、皮层、鞘细胞层和中心组织(侵染组织)。在中心组织的侵染细胞中,分布有大量的线粒体、高尔基体、核糖体及内质网,白车轴草根瘤侵染细胞中的细菌圆形或椭圆形,有明显的周膜、细菌细胞壁和质膜,在细菌发育过程中,周膜活动旺盛,有时相邻细菌的周膜发生融合,在周膜附近常分布有大量的内质网、高尔基体以及高尔基体小泡,似与周膜融合有关。紫花苜蓿细菌椭圆形、长棒形,甚至有的细菌呈分枝状。二者细菌细胞质中分布着大量的核糖体和纤维状的核物质。在白车轴草中还有染色很深的多聚磷酸盐颗粒。     

Involvement of polyamines in the drought resistance of rice

This study investigated whether and how polyamines (PAs) in rice (Oryza sativa L.) plants are involved in drought resistance. Six rice cultivars differing in drought resistance were used and subjected to well-watered and water-stressed treatments during their reproductive period. The activities of arginine decarboxylase, S-adenosyl-L-methionine decarboxylase, and spermidine (Spd) synthase in the leaves were significantly enhanced by water stress, in good agreement with the increase in putrescine (Put), Spd, and spermine (Spm) contents there. The increased contents of free Spd, free Spm, and insoluble-conjugated Put under water stress were significantly correlated with the yield maintenance ratio (the ratio of grain yield under water-stressed conditions to grain yield under well-watered conditions) of the cultivars. Free Put at an early stage of water stress positively, whereas at a later stage negatively, correlated with the yield maintenance ratio. No significant differences were observed in soluble-conjugated PAs and insoluble-conjugated Spd and Spm among the cultivars. Free PAs showed significant accumulation when leaf water potentials reached -0.51 MPa to -0.62 MPa for the drought-resistant cultivars and -0.70 MPa to -0.84 MPa for the drought-susceptible ones. The results suggest that rice has a large capacity to enhance PA biosynthesis in leaves in response to water stress. The role of PAs in plant defence to water stress varies with PA forms and stress stages. In adapting to drought it would be good for rice to have the physiological traits of higher levels of free Spd/free Spm and insoluble-conjugated Put, as well as early accumulation of free PAs, under water stress.     

Using the Pollen Viability and Morphology for Fluoride Pollution Biomonitoring

The methods using plants for biomonitoring of air and soil quality are simple, cheap, and fast and can supplement the classical physicochemical methods. In this study, biological pollen characterization of some collected legume species from an aluminum smelter area in Iran (IRALCO) was carried out to determine the actual value of pollen as a bioindicator of the effects of soil and atmospheric pollution. Young buds and flowers of six legumes (Cercis siliquastrum L., Medicago sativa L., Robinia pseudoacacia L., Melilotus officinalis (L.) lam, Trifolium repens L., and Sophora alopecuroides L.) in polluted and control plants were removed and compared. Studies of light and electron microscopic preparation showed some abnormalities during pollen development in affect of fluoride pollution. The viability of pollen grains estimated by staining with acetocarmine shows sharp differences in smearing advanced pollen grains from abnormal ones. Except M. officinalis, the pollen grains of C. siliquastrum, M. sativa, R. pseudoacacia, T. repens, and S. alopecuroides in polluted areas showed light, partial, or no staining with acetocarmine, whereas almost all of the control ones clearly stained. Observation of the pollen grains by light microscopy and scanning electron microscopy showed the significant effect of fluoride on shapes and sizes of pollen grains. The stimulation and inhibition of these pollen characteristics depend on the pollen species as well as on the pollutant and its concentration. Therefore, pollen grains provide essential information on biological impact of pollutants and they are good candidates for biomonitoring the atmospheric and edaphic pollutions.     

铜胁迫下链格孢菌对白车轴草生理生态的影响

通过盆栽实验研究了Cu胁迫下接种病原菌链格孢菌(Alternaria tenuis Nees)对白车轴草(Trifolium repens L.)生理生态指标的影响.结果表明:在未接种的对照组中,随着Cu浓度的增大(0~3000mg.kg-1),植株叶片失绿,生物量下降,叶绿素a、b、a b和类胡萝卜素及叶片可溶性蛋白质含量下降;植物体内丙二醛(MDA)高度积累,细胞膜结构遭到损坏,电导率增大;植株活性氧清除系统受损,不能保持原有平衡,超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性显著降低,过氧化物酶(POD)活性升高.接种链格孢菌后,植株叶片色素、膜系统及保护酶系统的损伤均加重,与不接种对照组相比,相同浓度Cu处理的叶绿素a、b、a b和类胡萝卜素、可溶性蛋白质含量、SOD和CAT活性均呈不同程度的下降,而电导率、MDA含量和POD活性则有所上升.     

光照强度和氮水平对白三叶幼苗生长与光合生理特性的影响

采用植物生长箱溶液培养方式,对白三叶幼苗进行了不同光强(2个水平)和氮浓度(5个水平)处理,探讨其生长、生物量和光合生理特征对生境变化的响应.结果表明:两种光强下白三叶幼苗茎和叶生物量随氮素浓度呈先升高后降低,而根系生物量和根冠比则随氮素浓度增高而降低.光照强度降低使白三叶幼苗根、茎、叶和整株生物量分别降低67.8%、29.9%、42.5%和45.2%;低光处理使幼苗的根冠比显著下降,而比叶面积(SLA)明显提高.幼苗根系体积随氮素浓度增高而降低,高生长光强根系体积显著高于低生长光强下的白三叶.幼苗根系表面积、根系长度和根系直径随氮素浓度增加呈先增加后降低趋势,两种不同生长光强下幼苗根系长度和根系直径差异显著,而根系表面积差异不明显.白三叶叶片光合速率(Pn)随氮素浓度增加呈先增加后降低趋势,高生长光强白三叶Pn显著高于低生长光强下的白三叶.两种生长光强间叶片气孔导度(Gs),胞间CO2浓度(Ci)和蒸腾速率(Tr)无显著差异,但氮素浓度对叶片Gs、Ci和Tr均有显著影响.光、氮及其交互作用对白三叶幼苗生长发育产生了显著影响,光照不足和氮缺乏都将导致白三叶幼苗生长减弱,但幼苗对这些不利环境具有较强的调节和适应能力.     

狗牙根、白三叶草对淡剑纹灰翅夜蛾生长发育和繁殖的影响

利用室内饲喂法,以高羊茅分别为对照,研究狗牙根(Cynodon dacyton)、白三叶草(Trifolium repens)对淡剑纹灰翅夜蛾Spodoptera depravata(Butler)存活、生长发育及繁殖的影响。结果表明,用狗牙根、白三叶草饲喂淡剑纹灰翅夜蛾的化蛹率、幼虫成蛾率均显著低于对照组。幼虫历期显著长于对照组。用狗牙根、白三叶草饲喂幼虫时,淡剑纹灰翅夜蛾的蛹期、产卵前期、产卵期、产卵量、♀成虫寿命与对照组均无显著差异,但幼虫龄期明显增加。狗牙根、白三叶草饲喂的淡剑纹灰翅夜蛾种群增长参数与对照组相比存在明显差异:(1)世代净生殖率(R0);(2)内禀增长率(rm);(3)周限增长率(λ);(4)种群数量趋势指数(I)明显降低;世代平均周期(T)和种群加倍时间(DT)明显延长。由此可见,狗牙根和白三叶草并不是淡剑纹灰翅夜蛾喜好的寄主植物。     

种间竞争梯度对白车轴草与其病原菌白车轴草单孢莠菌相互作用的影响

白车轴草（Trifolium repens)在与其病原菌白车轴草单孢锈菌（Uromyces trifolii-repentis)的长期相互作用中分化形成了抗病型（Resistance)无性系和易感型（Susceptibility)无性系。该研究工作旨在了解：1）在种间竞争不断增强的环境梯度中,抗型无性系和易感型无性系的生长表现有何区别？2）在同样的实验条件下,分别对抗病型无性系和易感型无性系进行接种感染后,两者的生长表现又有何区别？在一严重感病的白车轴草自然种群中,分别标定了17个抗病型无性系和14个易感型无性系,从各元性系的匍匐茎上分别剪取长5cm,具两个叶两个腋芽的小段作实验材料。接种孢子采自同一植物种群植株的病叶。实验在一个处在次生演替阶段的草本植物群落上进行,在该群落的种间竞争梯度上选定3个分别具有弱、中等和强种间竞争的生境（Habitat)建立实验站,每站设置等量的实验和对照组。结果显示：1）随着环境中种间竞争强度的增加,无论接种与否,抗病型和易感型白车轴草的叶生长量均凝减;2）在弱或强种间竞争环境条件下,无论接种与否,易感型无性系的生长均好于抗抗病型无性系;但在中等间竞争强度的环境条件下,对照实验中易感型无性系的生长显著好于抗病型无性系,而接种实验中的结果相反。这一研究结果表明,种间竞争强度可能是影响给定生境中寄主植物抗病型和易感型遗传型比率的重要因子之一,这一影响还将随着病原菌存在而否而发生改变。     

The host plant range of the pea aphid subspecies Acyrthosiphon pisum ssp. destructor (Johnson) (Hom., Aphididae)

Abstract:  In order to establish the host range of the pea aphid subspecies, Acyrthosiphon pisum ssp. destructor , and hence from which plant species pea crops are likely to become infested, the performance of this aphid on different leguminous plants was assessed. The plant species used were: Lotus uliginosus , Medicago sativa , Melilotus officinalis , Ononis repens , Sarothamnus scoparius , Trifolium hybridum , Trifolium pratense , Trifolium repens , Vicia cracca and Vicia faba . Vicia faba and Trifolium hybridum were the plants on which aphids reached the greatest size, took the least time to reach maturity, and experienced the lowest mortality. The time taken for the aphids to develop to maturity was negatively correlated with adult size, whereas survival to maturity was positively correlated with adult size. The host preference of the aphids was also assessed. The plant species selected as hosts by alatae were those on which their offspring performed best.